Research to develop instrumentatIon and methods for the sequence analysis[unreadable] of peptide antigens presented to the immune system in association with[unreadable] class I and class II molecules of the major histocompatibility complex is[unreadable] proposed. This is a necessary first step in the development of[unreadable] synthetic/recombinant vaccines or other modulators of the immune system[unreadable] that are effective against bacterial and viral infections, cancer,[unreadable] autoimmune disorders and tissue transplant rejection. Of highest priority[unreadable] is the development of methods for characterizing specific disease state[unreadable] associated antigens found in complex mixtures of several thousand self[unreadable] peptides. Multistage chromatography in conjunction with both[unreadable] microcapillary high performance liquid chromatography/ and high[unreadable] performance capillary electrophoresis/ electrospray ionization tandem mass[unreadable] spectrometry will be employed for this purpose. Both of the above systems[unreadable] will be coupled directly to cytotoxicity assays. Specific goals include[unreadable] the following: (a) to further improve the sensitivity of the tandem mass[unreadable] spectrometry method so as to facilitate sequence analysis of class I[unreadable] peptides at the low femtomole level, (b) to develop proteolytic digestion[unreadable] methods that will facilitate sequence analysis of class II antigens at the[unreadable] low femtomole level, (c) to sequence peptides presented by class I HLA-[unreadable] A2.l molecules on human melanoma cells that are recognized by melanoma[unreadable] specific cytotoxic T lymphocytes, (d) to sequence peptides presented by[unreadable] the human class I molecules, HLA-B7, Aw68, and Aw69, (e) to determine[unreadable] the structural basis of peptide interaction with class I, MHC molecules[unreadable] and to define peptide epitopes recognized by xenoreactive, HLA-A2.1[unreadable] restricted CTL, (f) to sequence peptides processed from HIV virus proteins[unreadable] and presented in association the class 1, HLA-A2.l molecules, (g) to[unreadable] identify the range of endogenous influenza hemagglutinin pep tides[unreadable] associated with the murine MHC class 1 Kd molecule produced by the wild[unreadable] type hemagglutinin gene and synthetic minigenes and to identify endogenous[unreadable] hemagglutinin peptides that bind Kd but are not immunogenic.[unreadable] [unreadable]